Antimutagenicity of secondary metabolites from higher plants.

Higher plants contain both mutagens and antimutagens and are susceptible to mutagenesis but screening programs for detection of antimutagenesis rarely employ higher plant systems. Short-term bacterial and mammalian tissue culture systems are the norm. Using modified screening tests for detecting antimutagenic agents, higher plants have been shown to contain a variety of structurally novel antimutagenic agents. Systematic bioassay-directed methodology resulted in the isolation in pure form and biological and chemical characterization of the responsible individual active components from various plants. The methodology in use is illustrated by the isolation of cinnamic acid, cinnamyl cinnamate and cinnamyl ricinoleate as the active constituents of the classic medicinal plant product, Styrax asiatica. The methods which may be used to reveal structure-activity relationships and to explore putative molecular modes of action are illustrated with excerpts from the same study.     

Naphthoquinone-lactones and extended quinones from Ventilago calyculata

New quinones have been isolated from the root bark of Ventilago calyculata. Ventilatones A and B are benzisochromanquinones, related to the ventiloquinones, which have an additional fused lactone ring while ventileins A and B are benzisochroman dimers having a dihydroxy-peri-xanthenoxanthenequinone chromophore.     

Benzisochromanquinones in ventilago species

From the acetone extract of the root bark of Ventilago maderaspatana eight new benzisochromanquinones; ventiloquinones A, B, C, D, E, F, G and H, have been isolated. Ventiloquinones I, J and K are three more new benzisochromanquinones isolated from the root bark of V. calyculata. The majority are 3,4,5,10-tetrahydro-cis-1,3-dimethyl-1H-naphtho[2,3-c]pyran-5,10-quinones related to eleutherin, but F, H, I, J and K are 6,9-quinones related to ventilagone.     

Naphthalenes and naphthoquinones from Ventilago species

Two new naphthalene derivatives and three naphthoquinones have been found in the root bark of Ventilago maderaspatana. Their structures are 2-acetyl-6,7-dimethoxy-3-methyl-1,8-methylenedioxynaphthalene (ventilaginone) and 1,3-dihydro-6,9-dihydroxy-7,8-dimethoxy-1-methylnaphtho[2,3-c]furan-3-one (ventilagol), 2(2′-acetoxypropyl)-3-hydroxy-5,7,8-trimethoxy-1,4-naphthoquinone (maderone), cordeauxione and isocordeauxione. The root bark of V. calyculata contains 2-methoxystypandrone and 1-hydroxy-6-methoxy-3-methylxanthone-8-carboxylic acid (calyxanthone).     

Probing structurally altered and aggregated states of therapeutically relevant proteins using GroEL coupled to bio-layer interferometry

The ability of a GroEL-based bio-layer interferometry (BLI) assay to detect structurally altered and/or aggregated species of pharmaceutically relevant proteins is demonstrated. Assay development included optimizing biotinylated-GroEL immobilization to streptavidin biosensors, combined with biophysical and activity measurements showing native and biotinylated GroEL are both stable and active. First, acidic fibroblast growth factor (FGF-1) was incubated under conditions known to promote (40°C) and inhibit (heparin addition) molten globule formation. Heat exposed (40°C) FGF-1 exhibited binding to GroEL-biosensors, which was significantly diminished in the presence of heparin. Second, a polyclonal human IgG solution containing 6–8% non-native dimer showed an increase in higher molecular weight aggregates upon heating by size exclusion chromatography (SEC). The poly IgG solution displayed binding to GroEL-biosensors initially with progressively increased binding upon heating. Enriched preparations of the IgG dimers or monomers showed significant binding to GroEL-biosensors. Finally, a thermally treated IgG1 monoclonal antibody (mAb) solution also demonstrated increased GroEL-biosensor binding, but with different kinetics. The bound complexes could be partially to fully dissociated after ATP addition (i.e., specific GroEL binding) depending on the protein, environmental stress, and the assay’s experimental conditions. Transmission electron microscopy (TEM) images of GroEL-mAb complexes, released from the biosensor, also confirmed interaction of bound complexes at the GroEL binding site with heat-stressed mAb. Results indicate that the GroEL-biosensor-BLI method can detect conformationally altered and/or early aggregation states of proteins, and may potentially be useful as a rapid, stability-indicating biosensor assay for monitoring the structural integrity and physical stability of therapeutic protein candidates.     

Role of antimutagens/anticarcinogens in cancer prevention

Recently it has become increasingly clear that chemicals found in our foods and beverages can prevent the genetic damage that leads to cancer initiation. Such substances may also affect subsequent events in the pathways that lead to cancer, and may have the potential to inhibit the mutations that allow tumor cells to become resistant to antitumor agents. We describe here the antimutagenic potential of Glabrene analogs against EMS-induced mutations utilizing modified Ames tests in S. typhimurium TA 100 and E. coli JC 5088. Results of studies of the ability of well-known antioxidants such as EGCG and related compounds to prevent drug resistance mutations in microorganisms are described, and their possible significance in the prevention of chemotherapeutic drug-resistance in tumor cells is discussed.     

Anthraquinones in Ventilago species

Eleven anthraquinones have been isolated from the root bark of Ventilago calyculata of which xanthorin-5-methyl ether and 2-hydroxyislandicin are n     

Flavonol 3-glycosides from the leaves of Flemingia stricta

A new glycoside, tamarixetin 3-rhamnoside together with kaempferol 3-rhamnoside, mearnsetin 3-rhamnoside, quercetin 3-rhamnoside, myricetin 3-rhamnoside and sitosterol glucoside, was identified from the leaves of Flemingia stricta     

Two anthraquinones from Ventilago calyculata

Two new anthraquinones have been isolated from the root bark of ventilago calyculata and their structures shown to be 2,4,8-trihydroxy-1-methoxy-3-methylanthraquinone and 2,4,8-trihydroxy-1,6-dimethoxy-3-methyl-anthraquinone.     

Antimicrobial agents from higher plants: Activity and structural revision of flemiflavanone-D from Flemingia stricta

Flemiflavanone-D from Flemingia stricta is active in vitro against Staphylococcus aureus and Mycobacterium smegmatis. Reexamination and interpretation of its spectral properties required revision of its molecular formula to C₂₅H₂₈O₆ and its structure to 2S-5,7,4′-trihydroxy-6-γ,γ-dimethylallyl-3′-γ,γ-dimethylallyl-oxidoflavan-4-one. The new structure was confirmed when reaction with chlorotrimethylsilane sodium iodide in acetonitrile deoxygenated and cyclized flemiflavanone-D to the known dicycloeuchrestaflavanone A. The absolute stereochemistry of flemiflavanone-D was established to be 2S by circular dichroism measurements.